Patterning of the Drosophila embryonic mesoderm requires the regulation of cell type-specific factors in response to dorsoventral and anteroposterior axis information. For the dorsoventral axis, the homeodomain gene, tinman, is a key patterning mediator for dorsal mesodermal fates like the heart. However, equivalent mediators for more ventral fates are unknown. We show that D-six4, which encodes a Six family transcription factor, is required for the appropriate development of most cell types deriving from the non-dorsal mesoderm - the fat body, somatic cells of the gonad, and a specific subset of somatic muscles. Misexpression analysis suggests that D-Six4 and its likely cofactor, Eyes absent, are sufficient to impose these fates on other mesodermal cells. At stage 10, the mesodermal expression patterns of D-six4 and tin are complementary, being restricted to the dorsal and non-dorsal regions respectively. Our data suggest that D-six4 is a key mesodermal patterning mediator at this stage that regulates a variety of cell-type-specific factors and hence plays an equivalent role to tin. At stage 9, however, D-six4 and tin are both expressed pan-mesodermally. At this stage, tin function is required for full D-six4 expression. This may explain the known requirement for tin in some non-dorsal cell types. (c) 2006 Elsevier Inc. All rights reserved.